Oral Iron Absorption of Ferric Citrate Hydrate and Hepcidin-25 in Hemodialysis Patients: A Prospective, Multicenter, Observational Riona-Oral Iron Absorption Trial.

Oral ferric citrate hydrate (FCH) is effective for iron deficiencies in hemodialysis patients; however, how iron balance in the body affects iron absorption in the intestinal tract remains unclear. This prospective observational study (Riona-Oral Iron Absorption Trial, R-OIAT, UMIN 000031406) was conducted at 42 hemodialysis centers in Japan, wherein 268 hemodialysis patients without inflammation were enrolled and treated with a fixed amount of FCH for 6 months. We assessed the predictive value of hepcidin-25 for iron absorption and iron shift between ferritin (FTN) and red blood cells (RBCs) following FCH therapy. Serum iron changes at 2 h (ΔFe2h) after FCH ingestion were evaluated as iron absorption. The primary outcome was the quantitative delineation of iron variables with respect to ΔFe2h, and the secondary outcome was the description of the predictors of the body's iron balance. Generalized estimating equations (GEEs) were used to identify the determinants of iron absorption during each phase of FCH treatment. ΔFe2h increased when hepcidin-25 and TSAT decreased (-0.459, -0.643 to -0.276, p = 0.000; -0.648, -1.099 to -0.197, p = 0.005, respectively) in GEEs. FTN increased when RBCs decreased (-1.392, -1.749 to -1.035, p = 0.000) and hepcidin-25 increased (0.297, 0.239 to 0.355, p = 0.000). Limiting erythropoiesis to maintain hemoglobin levels induces RBC reduction in hemodialysis patients, resulting in increased hepcidin-25 and FTN levels. Hepcidin-25 production may prompt an iron shift from RBC iron to FTN iron, inhibiting iron absorption even with continued FCH intake.

Evidence of accumulation of tetrodotoxin (TTX) in tissues and body parts of ectoparasitic copepods via their feeding on mucus of TTX-bearing pufferfish.

Adults of the ectoparasitic copepod Caligus fugu found on tetrodotoxin (TTX)-bearing pufferfish such as Takifugu alboplumbeus and Takifugu flavipterus are known to accumulate TTX in body tissues and parts other than the ovaries, oviducts, eggs, and cuticles. This study aimed to demonstrate, using immunoenzymatic staining techniques, that the TTX-free planktonic/infective copepodid stage of C. fugu could accumulate TTX in the tissues after molting into the parasitic stage (chalimus I) and then fed on mucus of host puffers. All the tissues of the planktonic copepodids were completely TTX-free, whereas chalimus I copepods accumulated TTX in parts other than the cuticles, guts, and some muscles. Chalimus IV and adult copepods retained TTX in these body parts but not in the reproductive organs, which were TTX-resistant, indicating that TTX was not vertically transmitted via eggs. Non-cellular TTX-positive contents found in the guts of some chalimi and adults indicated that the copepods potentially accumulated TTX by feeding on host mucus rather than skin tissues and blood. This study revealed that the presence or absence of TTX in some body parts differed among individuals of the parasite.

Evaluation of the tetrodotoxin uptake ability of pufferfish Takifugu rubripes tissues according to age using an in vitro tissue slice incubation method.

The tetrodotoxin (TTX) uptake ability of pufferfish Takifugu rubripes tissues and its growth-associated changes were investigated using an in vitro tissue slice incubation method. Tissue slices prepared from the liver, skin, and intestine of a non-toxic cultured adult T. rubripes (20 months old) and incubated with incubation buffer containing 25 µg/mL TTX for 1-48 h showed a time-dependent increase in the TTX content in all tissues. The TTX contents of the skin and intestine slices were comparable to or slightly higher than that of the liver slices, with a similar transition pattern, suggesting similar TTX uptake ability among the skin, intestine, and liver. The TTX uptake ability of the liver and intestine did not differ significantly between young (8 months old) and adult (20 months old) fish, but the skin slices of young fish took up approximately twice as much TTX as that of adult fish, suggesting that the TTX uptake ability of the skin is involved in the growth-dependent changes in the toxin distribution inside the body in T. rubripes. To estimate the TTX uptake pathway in each tissue, an immunohistochemical technique was used to observe temporal changes in the intra-tissue microdistribution of TTX during incubation. The findings suggested that TTX is transferred and accumulates from pancreatic exocrine cells to hepatic parenchymal cells in the liver, from connective tissues to basal cells in the skin, and from villi epithelial cells via the lamina propria to the muscle layer in the intestine.

A PQ-loop protein Ypq2 is involved in the exchange of arginine and histidine across the vacuolar membrane of Saccharomyces cerevisiae.

In nutrient-rich conditions, basic amino acids are actively accumulated into the vacuoles by H+-coupled transporters in Saccharomyces cerevisiae. In addition to the H+-coupled systems, the existence of an exchanger for arginine and histidine was indicated by kinetic analysis using isolated vacuolar membrane vesicles; however, the gene(s) involved in the activity has not been identified. Here, we show that the uptake activity of arginine driven by an artificially imposed histidine gradient decreased significantly by the disruption of the gene encoding vacuolar PQ-loop protein Ypq2, but not by those of Ypq1 and Ypq3. The exchange activity was restored by the expression of YPQ2. Furthermore, the substitution of a conserved proline residue, Pro29, in Ypq2 greatly decreased the exchange activity. These results suggest that Ypq2 is responsible for the exchange activity of arginine and histidine across the vacuolar membrane, and the conserved proline residue in the PQ-loop motif is required for the activity.

Physicochemical risk factors for building-related symptoms in air-conditioned office buildings: Ambient particles and combined exposure to indoor air pollutants.

We conducted a cross-sectional epidemiological study to examine the correlation between indoor air quality (IAQ) and building-related symptoms (BRSs) of office workers in air-conditioned office buildings. We investigated 11 offices during winter and 13 offices during summer in 17 buildings with air-conditioning systems in Tokyo, Osaka, and Fukuoka, and we included 107 office workers during winter and 207 office workers during summer. We conducted environmental sampling for evaluating IAQ and concurrently administered self-reported questionnaires to collect information regarding work-related symptoms. Multivariate analyses revealed that upper respiratory symptoms showed a significant correlation with increased indoor temperature [odds ratio (OR), 1.55; 95% confidence interval (CI), 1.11-2.18] and increased indoor concentration of suspended particles released from the ambient air pollution via air-conditioning systems (OR, 1.31; 95% CI, 1.08-1.59) during winter. In particular, smaller particles (particle size>0.3µm), which possibly penetrated through the filter media in air-conditioning systems from ambient air, were correlated with upper respiratory symptoms. The use of high-efficiency particulate air filters in air-conditioning systems and their adequate maintenance may be an urgent solution for reducing the indoor air concentration of submicron particles. Several irritating volatile organic compounds (VOCs) (e.g., formaldehyde, acetaldehyde, ethylbenzene, toluene, and xylenes) that were positively correlated with the indoor air concentration among their VOCs, were associated with upper respiratory symptoms, although their indoor air concentrations were lower than those specified by the indoor air quality guideline. A new approach and strategy for decreasing the potential combined health risks (i.e., additive effect of risks) associated with multiple low-level indoor pollutants that have similar hazardous properties are required.

Evaluating prevalence and risk factors of building-related symptoms among office workers: Seasonal characteristics of symptoms and psychosocial and physical environmental factors.

BACKGROUND: Psychosocial and environmental factors at the workplace play a significant role in building-related symptoms (BRSs). Environmental factors change during summer cooling and winter heating using air-conditioning systems. Thus, significant risk factors in each season need to be clarified. METHODS: A nationwide cross-sectional study was conducted during summer in Japan and seasonal differences between summer and winter were evaluated. Self-administered questionnaires were distributed to 489 offices. Possible risk factors for BRSs associated with the work environment, indoor air quality, and job stressors were examined by multiple regression analyses. RESULTS: Among people having at least one BRS, the prevalence of BRSs in summer (27.8%) was slightly higher than that in winter (24.9%). High prevalence was observed for eye and nasal symptoms related to dryness and general symptoms related to psychological distress in both seasons. Analyses revealed that dryness of air was an important and significant risk factor associated with BRSs, and job stressors were significantly associated with general symptoms in both seasons. Conversely, humidity was a significant risk factor of general symptoms in summer (odds ratio, 1.20; 95% confidence interval, 1.02-1.43). Carpeting, recently painted walls, and unpleasant chemical odors in summer and noise, dust and dirt, and unpleasant odors such as body or food odors in both seasons were significant risk factors for BRSs. CONCLUSIONS: Improvements in the physical environmental qualities in an office throughout the year are important along with the reduction in psychological distress related to work.

Ypq3p-dependent histidine uptake by the vacuolar membrane vesicles of Saccharomyces cerevisiae.

The vacuolar membrane proteins Ypq1p, Ypq2p, and Ypq3p of Saccharomyces cerevisiae are known as the members of the PQ-loop protein family. We found that the ATP-dependent uptake activities of arginine and histidine by the vacuolar membrane vesicles were decreased by ypq2Δ and ypq3Δ mutations, respectively. YPQ1 and AVT1, which are involved in the vacuolar uptake of lysine/arginine and histidine, respectively, were deleted in addition to ypq2Δ and ypq3Δ. The vacuolar membrane vesicles isolated from the resulting quadruple deletion mutant ypq1Δypq2Δypq3Δavt1Δ completely lost the uptake activity of basic amino acids, and that of histidine, but not lysine and arginine, was evidently enhanced by overexpressing YPQ3 in the mutant. These results suggest that Ypq3p is specifically involved in the vacuolar uptake of histidine in S. cerevisiae. The cellular level of Ypq3p-HA(3) was enhanced by depletion of histidine from culture medium, suggesting that it is regulated by the substrate.

Metabolic rate prediction in young and old men by heart rate, ambient temperature, weight and body fat percentage.

OBJECTIVES: An estimation of metabolic rate (MR) is needed to determine wet-bulb globe temperature (WBGT) reference values in order to reduce heat strain in physical workers. The aim of this study was to develop MR prediction equation for younger and older men in hot working environments. METHODS: We measured the MR and heart rate (HR) of both younger and older men at ambient temperatures (Ta) of 25, 30 and 35°C while they cycled on a bicycle ergometer at a workload of 30, 45 and 60% of maximal oxygen uptake (V̇O2max). Seven younger male university students aged 22.9 ± 0.7 (mean ± SD) years and seven older male workers aged 61.7 ± 2.2 (mean ± SD) years participated in this study. MR, HR and rectal temperature (Tre) were measured during the study. HR, ambient temperature (Ta), body weight (BW) and body fat percentage (BF) served as predictors of MR using multivariate analysis. To increase the MR prediction accuracy, the following three alternative predictors of HR were used: HRres, calculated as 100 × [(HR - resting HR) / (maximal HR - resting HR)]; HRnet, calculated as (HR - resting HR); and HRi, calculated as (HR / resting HR). RESULTS: The R(2) value indicated that the models with HRres or HRnet were more accurate than those with HRi or HR. Ta had a significantly positive correlation with MR in older men. BW had a significantly positive correlation with MR in both younger and older men, and BF had a significantly negative correlation with MR in both younger and older men. CONCLUSIONS: HRres or HRnet enabled more accurate MR prediction than HR. BW and BF would increase the accuracy of MR prediction.

Effects of water-damaged homes after flooding: health status of the residents and the environmental risk factors.

We evaluated the health status of residents and the environmental risk factors of housing after flooding. Questionnaires were distributed to 595 selected households (one adult resident per household) in six areas in Japan which were severely flooded between 2004 and 2010. A total of 379 responses were obtained. Indoor dampness and visible mold growth significantly increased in homes with greater flood damage. The incidence of respiratory, dermal, ocular, and nasal symptoms one week after flooding was significantly higher in flooded homes compared with non-flooded homes, the incidence of psychological disorders was significantly high for six months after flooding, and the incidence of post-traumatic stress disorder was significantly high six months after flooding. Significant risk factors for respiratory and nasal symptoms included proximity to industrial and waste incineration plants. Our results suggest that rapid action should be taken after flooding to ensure adequate public health and environmental hygiene in the water-damaged homes.

Spatial and temporal instability of local biotic community mediate a form of aposematic defense in newts, consisting of carotenoid-based coloration and tetrodotoxin.

Most animals advertise their unprofitability to potential predators via conspicuous signals. Whether the strength of this aposematic signal indicates the quality and quantity of chemical defenses in animals is controversial. Here, we investigated the relationship between the conspicuousness of an aposematic signal and toxicity, which likely depends, at least in part, on dietary sources, in the newt Cynops pyrrhogaster. Our results indicate that the magnitude of the aposematic signal was not correlated with the amount of tetrodotoxin (TTX) and 6-epi TTX of wild individuals among populations. Using atoxic newts, reared from eggs, we compared the ability to accumulate TTX from diets between mainland and island populations. Newts of a mainland population that exhibited a less conspicuous signal accumulated more TTX than did equivalent newts of an insular population that displayed a more conspicuous signal; this was unrelated to variation in the toxicity of wild individuals of these two populations. We also found toxicity of wild newts changed over approximately one generation (10 years) in both populations. These results indirectly suggest that environmental variance, such as fluctuations in TTX resources in nature, may obscure differences in the ability of wild newts to accumulate TTX, and that this variation may be responsible for a lack of correlation between the strength of a newt's signal and its toxicity in the wild. These results imply that toxicity of wild individuals likely is a phenotypic trait largely dependent on environmental conditions.

Comparison of the localization of tetrodotoxin between wild pufferfish Takifugu rubripes juveniles and hatchery-reared juveniles with tetrodotoxin administration.

To reveal the accumulation profile of tetrodotoxin (TTX) in pufferfish Takifugu rubripes juveniles, we compared the localization of TTX in various tissues among wild juveniles and hatchery-reared juveniles with or without TTX administration using immunohistochemical technique with anti-TTX monoclonal antibody. Immuno-positive reaction was observed in hepatic tissue, basal cell of skin and olfactory, olfactory epithelium, optic nerve and brain (optic tectum, cerebellum, medulla oblongata) of wild juveniles (body length: BL, 4.7-9.4 cm). TTX was detected in the same tissues as wild juveniles and epithelial cell layer of intestine of hatchery-reared juveniles (BL, 5.0-5.3 cm) to which TTX was orally administrated. No positive reaction was observed from the tissues of hatchery-reared juveniles without TTX administration. These results suggest that orally administrated TTX to the non-toxic cultured juveniles is accumulated in the same manner of wild juveniles. In addition, our study revealed that pufferfish accumulates TTX in the central nervous system.

Maturation-associated changes in the internal distribution of tetrodotoxin in the female goby Yongeichthys criniger.

Maturation-associated changes in the internal distribution of tetrodotoxin (TTX) in the goby Yongeichthys criniger were investigated in 29 and 40 female specimens collected from Okinawa, Japan, from August 2008 to June 2009 (Group I), and from November 2009 to August 2010 (Group II), respectively. In Group I, based on changes in the gonadosomatic index (GSI) and histologic observation of the ovary, the period from October 2008 through January 2009 was estimated to be the 'previtelline-forming period', February through March 2009 the 'vitelline-forming period', April through June 2009 the 'spawning period', and August 2008 the 'end of spawning period' of the preceding year. The TTX content (mouse unit [MU] per gram tissue) of each Y. criniger tissue (skin, muscle, liver, and ovary) quantified by liquid chromatography/mass spectrometry (LC/MS) was generally high during the spawning period and continued to rise until the end of spawning period, especially in the ovary. Total TTX per individual increased considerably during the spawning period, most of which located in the ovary, indicating that Y. criniger obtains a high amount of TTX during the spawning period, and accumulates most of it in the ovary. In contrast, the TTX content of the skin was highest at the end of spawning period, and most of the total TTX located in the skin during this period as well as during the previtelline-forming period. In Group II, the maturation stage of the ovaries of all specimens was determined, and the specimens were grouped accordingly. In the perinucleolus stage, yolk vesicle stage, and yolk globule stage I, most of the TTX was localized in the skin, but the TTX in the ovary greatly increased as the maturation stage advanced from yolk globule stage I to yolk globule stage III.

Transfer profile of orally and intramuscularly administered tetrodotoxin to artificial hybrid specimens of the pufferfish Takifugu rubripes and Takifugu porphyreus.

Tetrodotoxin (TTX) was administered to artificially hybridized specimens of the pufferfish Takifugu rubripes and Takifugu porphyreus to investigate toxin accumulation in hybrids and TTX transfer/accumulation profiles in the pufferfish body. In test fish administered TTX-containing feed homogenate at a dose of ∼400 MU/fish by oral gavage using a syringe (OGA group), the toxin content (MU/g tissue) of the digestive tract rapidly decreased and that of the liver increased from 1 to 24 h after administration. From 24 to 120 h, the toxin content of the liver decreased gradually, and the toxin appeared in the skin. On the other hand, intramuscularly administered TTX (400 MU/fish) was rapidly transferred to the liver and skin via the blood, and only a little toxin remained in the muscle even at 1 h (IMA group). The total amount of toxin remaining in the whole body (% of administered toxin) was 31-45% in the OGA group, and 42-74% in the IMA group; the scores in the OGA group were generally lower than those in the IMA group. In both OGA and IMA groups, the greatest amount of toxin accumulated in the liver (23-52%) after 8 h, followed by the skin (11-21%) after 72 h. The TTX administration experiment, especially using the oral gavage administration method, revealed that skins and livers of 'torama' pufferfish hybrid are endowed with TTX-accumulating ability, but the muscles are not, and that TTX taken up from toxic feed to the pufferfish body is transferred first to the liver and then to the skin via the blood.

Transfer profile of intramuscularly administered tetrodotoxin to artificial hybrid specimens of pufferfish, Takifugu rubripes and Takifugu niphobles.

Tetrodotoxin (TTX) was intramuscularly administered to artificially hybridized specimens of the pufferfish Takifugu rubripes and Takifugu niphobles to investigate toxin accumulation in hybrids, and TTX transfer/accumulation profiles in the pufferfish body. In the test fish administered 146 MU TTX in physiologic saline, TTX rapidly transferred from the muscle via the blood to other organs. Toxin transfer to the ovary rapidly increased to 53.5 MU/g tissue at the end of the 72-h test period. The TTX content in the liver and skin was, at most, around 4-6 MU/g tissue, and in the testis it was less than 0.01 MU/g tissue. On the other hand, based on the total amount of toxin per individual (% of the administered toxin), the skin and the liver contained higher amounts (20-54% and 2-24%, respectively), but the amount in the liver rapidly decreased after 8-12 h, and fell below the level in the ovary after 48 h. These findings suggest that part of the TTX is first taken up in the liver and then transferred/accumulated in the skin in male specimens and in the ovary in female specimens.

Tributyltin in blood of marine fish collected from a coastal area of northern Kyushu, Japan.

We investigated levels of the pollutant tributyltin (TBT) in blood of pufferfishes (six species), Japanese sea perch, red sea bream, Japanese common goby, Japanese flounder, rockfish, conger eel, and sea mullet collected off the coast of northern Kyushu, Japan. We found considerable levels of TBT (1.4-190 ng/mL) accumulated in the blood of these fish. Blood TBT concentrations were 1.3-22.5 times liver concentrations and 4.9-78 times muscle concentrations, except in conger eel and mullet. We detected TBT (16-111 ng/mL-blood) in the plasma of the fine-patterned puffer (Takifugupoecilonotus) year-round, without any apparent seasonal trend. These results suggest that fish inhabiting coastal areas of Kyushu, Japan, continue to be contaminated with TBT.

Maturation-associated changes in toxicity of the pufferfish Takifugu poecilonotus.

From October 2006 to December 2007, wild specimens of the pufferfish Takifugu poecilonotus (93 females, 45 males) were collected from the Ariake Sea. Tissue toxicity was examined by mouse bioassay, and tetrodotoxin (TTX) content in the blood plasma by enzyme-linked immunosorbent assay. The relationship between toxicity and maturation was investigated based on changes in the gonadosomatic index: December-March in females and November-March in males, the 'maturation period'; April, 'just after spawning'; and the other months, the 'ordinary period'. Toxicity of both sexes was high throughout the year, but sharply declined in April. In all tissues examined (skin, liver, and ovary) other than testis, toxicity exceeded 1000 MU/g or 10,000 MU/individual in many individuals. Seasonal profiles of tissue toxicity differed markedly between sexes. In females, liver toxicity was high during the ordinary period, and ovary toxicity was high during the maturation period. In males, little maturation-associated change in the toxin distribution was observed. Plasma TTX levels were similar between the sexes (1.59-15.1 MU/ml), and fluctuated largely throughout the year without corresponding changes in tissue toxicity. The percentage of TTX binding to high molecular-weight substances in the plasma varied in association with maturation; the binding ratio fluctuated at relatively low levels during the ordinary period, and stabilized at a high level during the maturation period.

Transfer profile of intramuscularly administered tetrodotoxin to non-toxic cultured specimens of the pufferfish Takifugu rubripes.

Tetrodotoxin (TTX) was intramuscularly administered to non-toxic cultured specimens of the pufferfish Takifugu rubripes to investigate TTX transfer/accumulation profiles in the pufferfish body. In two groups of test fish administered either 50MU/individual of TTX standard (purified TTX; PTTX) or crude extract of toxic pufferfish ovary (crude TTX; CTTX), TTX rapidly transferred from the muscle via the blood to other organs. The toxin transfer profiles differed between groups, however, from 4 to 72h. In the PTTX group, little TTX was retained in the liver, and most (>96%) of the toxin remaining in the body transferred/accumulated in the skin after 12h, whereas in the CTTX group, a considerable amount of toxin (15%-23% of the administered toxin or 28%-58% of the remaining toxin) was transferred/retained in the liver for up to 24h, despite the fact that 89% of the remaining toxin transferred/accumulated in the skin at the end of rearing period (168h). The total amount of toxin remaining in the entire body at 1-4h was approximately 60% of the administered toxin in both groups, which decreased at 8-12h, and then increased again to approximately 60%-80% at 24-168h. Immunohistochemical observation revealed that the toxin accumulated in the skin was localized at the basal cells of the epidermal layer.

The risk screening for indoor air pollution chemicals in Japan.

In recent years, public health problems caused by indoor air pollution have been drawing strong public concern in Japan. After conducting extensive exposure assessment, governmental agencies have taken effective measures to solve the problem; for instance, "Guidelines for indoor air quality (IAQ)" of 13 chemicals, for example, formaldehyde, toluene, and xylene, has been established. Thousands of chemicals have been identified in the indoor environment. Priority rating of those chemicals, however, was not based on the health risk level. We developed a risk-screening scheme for indoor air pollution chemicals and analyzed the current status of the risk levels of those chemicals in Japan. We researched scientific knowledge of health hazards and exposure surveys of indoor air pollution chemicals in Japan, and classified those chemicals based on the health risk level estimated from the scheme. The risk levels of 93 chemicals were characterized and six chemicals (formaldehyde, acrolein, 1,4-dichlorobenzene, benzene, tetrachloroethylene, and benzo(a)pyrene) were classified in the highest risk category.

Accumulation of tetrodotoxin (TTX) in Pseudocaligus fugu, a parasitic copepod from panther puffer Takifugu pardalis, but without vertical transmission--using an immunoenzymatic technique.

The caligid copepod Pseudocaligus fugu, a common parasite on the body surface of both toxic (Takifugu pardalis) and cultured, non-toxic (Takifugu rubripes) puffer fishes, was isolated and analyzed for the presence of tetrodotoxin (TTX) using a monoclonal anti-TTX antibody-based immunohistochemical technique. Histological sections of female P. fugu from Takifugu pardalis explicitly revealed that positive brown staining was visible in regions of gut and appendages, and also in the general body tissues from the prosome to the urosome. It is absent from the epicuticle, ovary, oviduct, uterus and egg sacs, where clear negative blue color reaction was obtained. In contrast, the caligids on cultured, non-toxic Takifugu rubripes, considered as negative control, had no sign of TTX. The results indicate that there is no vertical transmission of TTX in the parasitic caligids, which could acquire TTX by feeding on the toxic mucus and skin tissues of host puffer fish.